tbuckley/cabot-search huggingface dataset:
Abstract P2083: Erk-mediated Phosphorylation Of Grk2 Leads To Reduced Pyruvate Dehydrogenase Activity And Alters Pyroptotic Signaling
Authors: Ruxu Zhai, Sarah Montgomery, Tian Yuzhen, Emad Alnemri, Hsin Yao tang, Priscila Sato
Journal: Circulation Research (2022)

G protein-coupled receptors (GPCRs) are important regulators of cellular functions where agonist binding leads to receptor conformational changes and downstream activation of g-protein-mediated signaling cascades. Signal transduction is terminated by receptor phosphorylation mediated by G-protein coupled receptor kinases (GRKs). GPCR kinase 2 (GRK2) is a main GRK in the heart and is upregulated in heart failure (HF) patients. Recently, we and others have shown that GRK2 can translocate to cardiac mitochondria where it regulates metabolism. Notably, we found that phosphorylated GRK2 at S670 post- ischemia/reperfusion (IR) decreases glucose mitochondrial utilization and pyruvate dehydrogenase (PDH) activity. Furthermore, 2D-SDS PAGE followed by LC/MS/MS revealed a potential link between mitochondrial GRK2 and PDH phosphorylation. Using a novel PDHα KI CRISPR Cas HEK cell line, we tested the hypothesize that PDH phosphorylation of this novel site was important for PDH activity. Utilizing cytosolic and mitochondrially-targeted pyruvate indicators we measured pyruvate levels in WT and KI HEK cells. In parallel, using a novel mouse model GRK2-S670A, we investigated the role of ERK in mediating GRK2 mitochondrial translocation and pyroptotic signaling. Further experiments will determine long-term impact of mitochondrial GRK2 in cell survival in cardiac pathological conditions.
Abstract P2020: A Novel Molecular Mechanism Of Cardiac Hypertrophy
Authors: Yongluan Lin, Lei Lu, Liling Zheng, Shuping Zhong
Journal: Circulation Research (2022)

Proliferation of cardiac myocytes is accompanied by increases in cell mass to maintain myocyte size in fetal heart. In adult cardiac myocytes, cell growth becomes uncoupled from proliferation after growth stimuli, resulting in hypertrophic growth. Both proliferation and hypertrophy growth of cardiac myocytes require more protein synthesis. Brf1 (TF IIB-related factor 1) and Pol III genes (RNA Polymerase III-dependent genes) are the dominant factors of protein produce. Brf1 is a key regulator for Pol III genes. Cardiac hypertrophy is a critical risk factor for cardiovascular disease, including heart failure, arrhythmia, and sudden death. Cardiac hypertrophy involves both the processes of embryonic gene expression and transcriptional reprogramming. The processes are tightly modulated by different signal pathways and epigenetic regulation. Diabetic cardiomyopathy is associated with an increased risk for heart failure and death in patients with diabetes. To explore the molecular mechanism of cardiac hypertrophy, we investigate the alterations of Brf1 and Pol III gene transcription under glucose treatment by using cell culture model and animal model. We have found that glucose increases Brf1 expression and Pol III gene transcription. We identified that these genes are mediated by JNK1, MSK1, pAMPK and phosphorylated histone H3. inhibiting the pathways decrease Brf1 expression, reduce Pol III gene transcription, consequently repression of myocytes growth. It implies that developing an effective inhibitor to repress Brf1 expression may provide a potential approach for the therapy of cardiac hypertrophy. More studies on cardiac hypertrophy are going in our laboratory.
Abstract P2080: Role Of Atg9 Proteins In Regulating Autophagosome Formation
Authors: Rachel Y Diao, Wenjing Liang, Liguo Chi, Xi Fang, Asa Gustafsson
Journal: Circulation Research (2022)

Cardiac myocytes require large amounts of energy generated by mitochondria to sustain contraction, and there is a strong link between mitochondrial dysfunction and heart disease. Defective mitochondria are less efficient at generating ATP, produce excessive reactive oxygen species, and release pro-apoptotic proteins, all of which can lead to loss of cardiac myocytes and reduced ability to sustain contractile function. Therefore, mitochondrial quality control is critical in preventing mitochondria from causing harm to the cell. Selective autophagy of the mitochondria (mitophagy) is the primary mechanism responsible for eliminating dysfunctional mitochondria. Despite intense research focus on mitophagy over the past decade, little is still known about early events in the pathway, and the signals that orchestrate the selective recruitment of the autophagic machinery to dysfunctional mitochondria are unclear. It is known that autophagosome formation requires Atg9, which is embedded in small Golgi-derived vesicles. To date, most data on Atg9 are from yeast and our knowledge of mammalian Atg9a and Atg9b functions in cells and tissues are still limited. Here, we found that Atg9a and Atg9b are expressed in the heart and that both proteins are increased in the infarct border zone after ligation of the LAD. We also discovered that Atg9a and Atg9b are present on distinct vesicles in myocytes. Atg9b-positive vesicles are localized throughout the cytosol, whereas Atg9a-positive vesicles are associated with the mitochondria even under basal conditions. This suggests that they might have distinct functions and that Atg9a may play a more selective role at the mitochondria. Interestingly, Atg9a, but not Atg9b, is degraded by the proteasome upon activation of mitophagy, a potential indication that this may be a mechanism to prevent excessive mitophagy. Overall, our findings suggest that Atg9a and Atg9b play distinct roles in autophagy and mitophagy and that the mitochondrial localization of Atg9a allows it to initiate autophagosome formation more efficiently during mitophagy.
Abstract EC106: Cardiac Fibroblast GSK-3α Aggravates Ischemic Cardiac Injury By Promoting Fibrosis, Inflammation, And Impairing Angiogenesis
Authors: Prachi Umbarkar, Sultan Tousif, Suma Ejantkar, Angelica Toro Cora, Qinkun Zhang, Hind Lal
Journal: Circulation Research (2022)

Background: Myocardial Infarction (MI) is the leading cause of death worldwide. Numerous studies from our lab and others have implicated Glycogen Synthase Kinase-3 (GSK-3) as a promising therapeutic target for cardiovascular diseases. GSK-3 is a family of ubiquitously expressed serine/threonine kinases. GSK-3 isoforms appear to play overlapping, unique, and even opposing functions in the heart. Previously our group has identified that cardiac fibroblast (CF) GSK-3β acts as a negative regulator of fibrotic remodeling in the ischemic heart. However, the role of CF-GSK-3α in MI-induced adverse cardiac remodeling is not defined. Methods and Results: To determine the role of CF-GSK-3α in MI-induced adverse cardiac remodeling, GSK-3α was deleted specifically in the activated fibroblast/myofibroblast using tamoxifen (TAM)-inducible Periostin promoter-driven Cre recombinase system. At 12 weeks of age, mice were fed with TAM diet. After 1 week of TAM treatment, control (GSK-3α fl/fl Cre -/- ) and KO (GSK-3α fl/fl Cre +/- ) mice were subjected to permanent LAD ligation (MI) surgery. Echocardiographic analysis revealed that control mice developed severe systolic dysfunction and dilative cardiac remodeling post-MI. These MI-induced pathologies were remarkably prevented by myofibroblast-specific GSK-3α deletion. To further confirm the role of CF-GSK-3α in processes contributing to cardiac remodeling, we harvested the hearts at 4 weeks post-MI and analyzed signature genes of adverse remodeling. Specifically, qPCR analysis was performed to examine the gene panels of inflammation (TNFα, IL-6, IL-1β), fibrosis (COL1A1, COL3A1, COMP), and angiogenesis (VEGFR2, CD31, HIF-1α). As expected, ischemic injury induced the expression of inflammation and fibrosis-related genes in the control group. In contrast, CF-GSK-3α KO hearts did not display any of these hallmarks. Another critical feature of adverse cardiac remodeling is impaired angiogenesis. Indeed, the CF-GSK-3α KO hearts were protected from MI-induced reduction in angiogenesis-related genes. Taken together, our findings support a driving role of CF-GSK-3α in MI-induced adverse cardiac remodeling. Conclusion: Our findings suggest a critical role of CF-GSK-3α in pathological cardiac remodeling and heart failure that could be therapeutically targeted for future clinical applications.
Abstract P3101: Chromatin Remodeling BAF (SWI/SNF) Complex Controls Both Cardiogenesis And Neurogenesis From Human Pluripotent Stem Cells
Authors: Juli Liu, Sheng Liu, Jun Wan, Lei Yang
Journal: Circulation Research (2022)

Background: Early human heart and brain development simultaneously occur during embryogenesis. Notably, in human newborns, congenital heart defects strongly associate with neurodevelopmental abnormalities, suggesting a common gene or complex underlying both cardiogenesis and neurogenesis. However, due to lack of in vivo studies, the molecular mechanisms that govern both early human heart and brain development remain elusive. Results: Here, we report ARID1A, a DNA-binding subunit of the BAF(SWI/SNF) epigenetic complex, controls both neurogenesis and cardiogenesis from human embryonic stem cells (hESCs) through distinct mechanisms. Loss-of-ARID1A (ARID1A –/– ) leads to spontaneous differentiation of neural cells together with globally enhanced expression of neurogenic genes in undifferentiated hESCs. Additionally, when compared with WT hESCs, cardiac differentiation from ARID1A –/– hESCs is prominently suppressed, whereas neural differentiation is significantly promoted. Whole genome-wide scRNA-seq, ATAC-seq, and ChIP-seq analyses reveal that ARID1A is required to open chromatin accessibility on promoters of essential cardiogenic genes, and temporally associated with key cardiogenic transcriptional factors T and MEF2C during early cardiac development. However, during early neural development, transcription of most essential neurogenic genes is dependent on ARID1A, which can interact with a known neural restrictive silencer factor REST/NRSF. Conclusions: We uncover the key and opposite roles by the BAF(SWI/SNF) complex to govern early cardiac and neural development. Global chromatin accessibility on cardiogenic genes is dependent on ARID1A, whereas transcriptional activity of neurogenic genes is under control by ARID1A-REST/NRSF interaction.
Abstract EC102: RBPMS, An RNA-binding Protein That Mediates Cardiomyocyte Binucleation And Cardiovascular Development
Authors: Peiheng Gan, Rhonda S Bassel-Duby, Ning Liu, Eric N Olson
Journal: Circulation Research (2022)

Noncompaction cardiomyopathy is a common congenital cardiac disorder associated with abnormal ventricular cardiomyocyte trabeculation and impaired pump function. The genetic basis and underlying mechanisms of this disorder remain elusive. We show that genetic deletion of RNA binding protein with multiple splicing (Rbpms), an uncharacterized RNA binding and splicing factor, causes perinatal lethality in mice due to congenital cardiovascular defects, including noncompaction cardiomyopathy, ventricular septal defects, and patent ductus arteriosus. Loss of Rbpms causes premature onset of cardiomyocyte binucleation and cell cycle arrest during development, thus disturbing embryonic heart growth. Human iPSC-derived cardiomyocytes with RBPMS gene deletion have a similar blockade to cytokinesis, and Rbpms expression is decreased in human patients with congenital cardiomyopathy, implicating an essential role of Rbpms in human heart development. Paired-end RNA sequencing analysis revealed that RBPMS plays a role in RNA splicing and influences RNAs involved in cytoskeletal signaling pathways. We found that RBPMS mediates isoform switching of the heart-specific LIM domain protein Pdlim5. Loss of Rbpms leads to abnormal accumulation of Pdlim5-short isoforms, disrupting cardiomyocyte cytokinesis. Our findings connect premature cardiomyocyte binucleation to noncompaction cardiomyopathy and highlight the role of Rbpms in this process.
Abstract P3008: Endothelial Stearoyl-CoA Desaturase-1 Catalyzes Fatty Acid Metabolites To Mitigate Arterial Inflammation And Stiffness
Authors: Susana Cavallero, Mehrdad Roustaei, Ana M Blazquez-Medela, Sandro Satta, Henry Phan, Kyung In Baek, Rongsong Li, Tzung K Hsiai
Journal: Circulation Research (2022)

Introduction: Vascular metabolic plasticity is intimately linked with endothelial homeostasis. Stearoyl-CoA desaturase-1 (SCD1), the enzyme that catalyzes the saturated long-chain fatty acids (FA) into monounsaturated FA (MUFA), is responsive to pulsatile shear stress (PSS), and is associated with cancer cell metabolism. We assessed whether exercise-mediated PSS activates PPARγ-SCD1 to mediate anti-inflammatory metabolites for vascular protection. Methods and Results: Human aortic endothelial cells (HAEC) were subjected to PSS (τ avg = 23±8 dyne/cm 2 at 1 Hz) for 4 hours, followed by metabolomic analysis. SCD1-mediated fatty acid metabolites, azelaic acid (AZA) and oleic acid (OA), were identified to mitigate NFκB luciferase activity and monocyte binding ( p &lt; 0.05, n= 5, vs. static condition). PSS upregulated PPARγ and SCD1 mRNA expression, whereas knockdown of PPARγ using siRNA downregulated SCD1. Wild type mice (C57BL/6J) undergoing 2-wk voluntary exercise developed increased perinuclear SCD1 expression in the aortic endothelium ( p &lt; 0.05, n=6, vs. sedentary controls), whereas SCD1 expression was reduced in PPARγ -dominant negative mice following exercise. LDLR -/- mice with endothelial-specific SCD1 deletion (SCD1 EC-KO : Cdh5 Cre , Scd1 f/f ) on 4-wk high-fat diet (HFD) developed significantly increased VCAM-1 and Oil Red O positive lesions in the aortic arch, whereas exercise or selective overexpression (OE) of SCD1 in endothelial cells by pAd5/F35/CD144 adenoviral vector mitigated the lesions ( p &lt; 0.05, n=5, HFD + exercise or HFD+Ad-SCD1 vs. HFD). Arterial stiffness as measured by Pulse Wave Velocity of the left common carotid was elevated in LDLR -/- SCD1 EC-KO on HF diet, but was decreased with exercise or SCD1 OE. Conclusion: PPARγ-SCD1 signaling is flow-responsive to mitigate arterial inflammation and stiffness. Despite SCD1-associated cancer cell metabolism, endothelial-specific SCD1 confers vascular protection, consistent with the reported skeletal muscle-specific model. Thus, we demonstrate that exercise activates PPARγ-SCD1 and SCD1 catalyzes AZA and OA as biomarkers with therapeutic potential.
Adherence to entry criteria and one year experience of long-term oxygen therapy in Poland
Authors: D Gorecka, P Sliwinski, J Zielinski
Journal: European Respiratory Journal (1992)

The adherence to entry criteria and results of one year long-term oxygen therapy (LTOT) in Poland were analysed. Four hundred and seven patients with advanced respiratory failure due to chronic lung diseases qualified for LTOT were observed for one year in 12 regional LTOT centres. There were 315 patients with chronic obstructive pulmonary disease (COPD) and 92 with other chronic lung diseases. In 270 patients the prescription of oxygen was based on the single criterion of stable arterial oxygen tension (PaO2) less than or equal to 55 mmHg (7.3 kPa); and in the remaining 137 with less severe stable hypoxaemia (PaO2 56-65 mmHg) (7.4-8.6 kPa), on concomitant signs of cor pulmonale and/or of tissue hypoxia. Of 407 patients who started LTOT, 95 (23%) died during the first year of treatment. The mortality rate was 21% for COPD patients and 33% for patients with other lung diseases. After one year of LTOT 312 patients survived: 250 COPD patients and 62 with other lung diseases. Of these, 19 COPD patients (8%) and 9 with other chronic lung disease (15%) who had PaO2 less than or equal to 55 mmHg (7.3 kPa) when oxygen was prescribed, presented with PaO2 greater than 55 mmHg (7.3 kPa). From the 137 patients (106 with COPD and 31 with other lung diseases) who qualified with less severe hypoxaemia, 10 COPD patients (10%) and 5 (16%) with other lung disease had PaO2 greater than 65 mmHg (8.6 kPa).(ABSTRACT TRUNCATED AT 250 WORDS)
Abstract P3125: Cldn11 Is A Candidate For Vascular Smooth Muscle Cell Plasticity Regulator In Atherosclerosis Pathology
Authors: Hyung-Seok Kim, Su-Jin Lee, Soojin Kim, Jungho Lee, Jong-Tae Park
Journal: Circulation Research (2022)

Claudin-11(CLDN11) is a component of tight junction, and important for normal CNS functions. The function of CLDN11 is well elucidated in oligodendrocytes and CNS system, but its expression pattern according to human vascular bed such as intracranial and extracranial, role in the vascular smooth muscles (vSMCs) plasticity and atherosclerosis is still unknown. Human left anterior descending coronary artery (LAD) and middle cerebral artery (MCA) vascular smooth muscle layer was dissected from the dead body. CLDN11 expression is identified in LAD and MCA vascular smooth muscle cell layer, and the CLDN11 expression is higher in MCA than in LAD (p&lt;0.01). Interestingly, the CLDN11 expression is decreased in vSMCs around the atherosclerotic lesions compare to non-atheromatous lesions by immunohistochemistry. To determine the functional role of CLDN11 in vSMCs, microarray analysis was performed following siCLDN11 knockdown. Compared to normal HASMC (human aortic vascular smooth muscle cells), siCLDN11-HASMC was showed increased gene expression such as RSPO3, CXCL8, EGR3, CYP1B1, TNFAIP2, PTGS2, HEY1, BMPER and these set was related to cell migration, proliferation and angiogenesis. CLDN11 knockdown vSMCs showed increased the cell migration of HASMC measured by wound healing, tube formation, and cell cycle assay, and reduced cas-3 cleavage. Finally, CLDN11 expression was inversely correlated with the progression of coronary atherosclerosis and the sudden cardiac death with complicated coronary atherosclerosis. This study shows that CLDN11 play a role in the initiation and propagation of atherosclerosis by regulating the plasticity of vSMCs, suggesting the possibility of biological marker for atherosclerosis in clinical diagnosis (This work was supported by NRF-2019R1A2C1089108, NRF-2019M3E5D1A02068082, COMPA-2022C100, and BK21 Program, Center for Glocal Future Biomedical Scientists at Chonnam National University).
Abstract P2102: EB3 Inhibitor VT-109 Activated FOXM1 Pathways To Promote Resolution Of Acute Lung Injury
Authors: Man Long Kwok, Xinyan Qu, Wan Ching Chan, Shuangping Zhao, Jonathan Le, Mark Maienschein-Cline, Yulia A Komarova
Journal: Circulation Research (2022)

Acute respiratory distress syndrome (ARDS) is the acute onset of non-cardiogenic pulmonary edema, hypoxemia, and respiratory failure, conditions associated with a high mortality rate. ARDS is frequently caused by lung infections including SARS-CoV-2. In light of the COVID-19 pandemic, development of therapeutic approaches to combat ARDS have become an urgent unmet medical need. As increased vascular leakage is a culprit of alveolar damage and lung inflammation, we hypothesize that an effective therapy against vascular leakage should improve the clinical outcome of ARDS patients. We have identified a drug candidate Vascular Therapeutics (VT)-109, an allosteric inhibitor of End Binding protein 3, which contributes to vascular leakage by amplifying pathological calcium signaling in endothelial cells. Treatment of mice with VT-109 reduced the leakage of proteinous fluids in lungs to 55.4% (n=7, p&lt;0.05) after endotoxin challenge, 77.7% (n=6, p&lt;0.01) after exposure to high volume mechanical ventilation, and 42.5% (n=7, p&lt;0.05) in the two-hit model, a combination of polymicrobial sepsis with normal tidal volume mechanical ventilation. VT-109 reduced the mortality of both septic (n=10, p&lt;0.01) and endotoxin-challenged (n=10, p&lt;0.05) mice. Furthermore, treatment with VT-109 in mice expressing human ACE2 receptor blocked diffuse alveolar damage (n=5, p&lt;0.0001) caused by SARS-CoV-2 infection. In this COVID-19 model, the therapeutic benefits of VT-109 were observed when treatment was initiated at either day 1 or day 3 post-viral infection. Transcriptome analysis of lung endothelial cells in mice challenged with endotoxin showed that VT-109 upregulated 23 FOXM1 target genes (&gt;2-fold) suggesting that it promotes endothelial cell regeneration by activating the FOXM1 pathway. Our data demonstrate that VT-109 successfully treats vascular leakage and is a promising drug candidate for future clinical studies. Funding for this work is provided by the Congressionally Directed Medical Research Programs through the Peer Reviewed Medical Research Program (PRMRP) under award No. W81XWH-21-1-0639. Opinions, interpretations, conclusions, and recommendations are those of the author and are not necessarily endorsed by the Department of Defense.
Abstract P2058: LncRNA PCAT19 Limits Angiogenesis And Safeguards Quiescent Endothelial DNA By Preventing Uncontrolled Phosphorylation Of Replication Protein A2
Authors: James A Oo, Katalin Pálfi, Timothy Warwick, Ilka Wittig, Frederike Boos, Matthias Leisegang, Ralf P Brandes
Journal: Circulation Research (2022)

Background: Long noncoding RNAs (lncRNA) provide an additional layer of molecular control in pathways crucial for vascular development and cardiovascular disease. We set out to identify novel endothelial lncRNAs that could be exploited to treat vascular disease. Methods and Results: We identified the endothelial cell-specific long noncoding RNA (lncRNA) PCAT19, which contributes to the endothelial proliferation-quiescence switch and acts as a safeguard for the endothelial genome during quiescence. PCAT19 was upregulated with endothelial cell confluence and quiescence. Knockdown of PCAT19 promoted endothelial proliferation and angiogenic sprouting. Conversely, PCAT19 overexpression reduced endothelial proliferative capacity and angiogenic sprouting. Using a cardiac organoid model system, we observed a markedly denser vascular network after the removal of PCAT19. RNA-sequencing after PCAT19 knockdown revealed multiple differentially regulated cell cycle genes. An antisense-oligonucleotide pulldown of PCAT19 followed by mass spectrometry identified multiple DNA damage response and cell cycle-related proteins as PCAT19 interaction partners; among them was the full DNA replication protein A complex. PCAT19 knockdown was found to sensitise DNA to damage, as measured by heightened p53 and γH2AX levels as well as a positive TUNEL signal and longer comet tail olive moments. Mechanistically, PCAT19 limited the phosphorylation of RPA2 on the serine 33 (S33) residue by inhibiting RPA2 interaction with the ATR kinase. Conclusions: In healthy vessels, endothelial cells maintain a stable, differentiated and growth-arrested phenotype. Upon injury, a rapid phenotypic switch facilitates proliferation to restore tissue perfusion. PCAT19 was identified as a highly enriched endothelial lncRNA that acts as a dynamic guardian of the endothelial genome, promotes cellular longevity and facilitates rapid switching to proliferation. Therapeutic targeting of PCAT19 could potentially alter the course of vascular disease development and progression by modulating endothelial growth and angiogenic sprouting.